Inhibitory Effect Of Methionine Enkephalin On Growth Of Human Cervical Immortal H8 Cells

Objective:Squamous Intraepithelial Lesion（SIL）is a common disease of women of childbearing age,and its incidence is increasing worldwide in recent years.According to the cervical squamous intraepithelial lesion diagnosis and treatment guidelines,Low-grade Squamous Intraepithelial Lesion can be selected for follow-up or surgical treatment.For High-grade Squamous Intraepithelial Lesion,surgery is the main treatment method.However,surgery can cause various problems,such as postoperative abortion and psychological stress.If there is a drug that can cure cervical squamous intraepithelial lesions,it will greatly reduce the trauma to the patient.The cervical immortalized epithelial H8 cells（hereinafter referred to as H8 cells）is the immoralized cervical squamous cell line of HPV16,which is equivalent to clinical cervical intraepithelial neoplasia（similar to CINII-III）.The purpose of this article is to study the inhibitory effect of Methionine-enkephalin（MENK）on the growth of H8 cells.Method:Cervical immortalized H8 cells were cultured in vitro and treated with different concentrations of MENK for different periods of time.The cell activity was evaluated by MTS analysis and flow cytometry,the best action concentration and action time was selected,and morphological changes were observed under conventional optical microscope.Cell cycle changes were determined by flow cytometry after MENK treated H8 cells.MRNA expression level of Opioid growth factor receptor（OGFr）was determined by RT-qPCR.The apoptosis of H8 cells after MENK was determined by flow cytometry.Enzyme linked immunosorbent assay（Elisa）was used to determine the content of Transforming growth factor-alpha（TGF-）in H8 cells before and after drug administration.Immunofluorescence assay was used to determine the intracellular location of OGFr and its expression after drug administration.Result:MENK inhibits H8 cells by OGFr.MENK can lead to cell cycle arrest in G0/G1 phase,decrease the percentage of cells in S and G2/M phase,and H8 cell apoptosis was significantly promoted.Reverse transcription-quantitative polymerase chain reaction demonstrated that H8 cells increased opioid receptor expression by MENK treatment.In addition,the level of TGF-β₁expression in the MENK group was higher than the control group after treatment with MENK at different times.Conclusion:The results of this study demonstrate for the first time that MENK can inhibit the growth of H8 cells and induce apoptosis of H8 cells,and its underlying mechanism may be related to the expression of OGFr receptor and TGF-β₁.MENK may be an effective drug for the treatment of cervical intraepithelial neoplasia.